Homologous recombination-mediated gene targeting in the liverwort Marchantia polymorpha L

The liverwort Marchantia polymorpha is an emerging model organism on account of its ideal characteristics for molecular genetics in addition to occupying a crucial position in the evolution of land plants. Here we describe a method for gene targeting by applying a positive/negative selection system...

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Bibliographic Details
Published in:Scientific reports 2013-03, Vol.3 (1), p.1532-1532, Article 1532
Main Authors: Ishizaki, Kimitsune, Johzuka-Hisatomi, Yasuyo, Ishida, Sakiko, Iida, Shigeru, Kohchi, Takayuki
Format: Article
Language:English
Subjects:
631/1647/1513/1967
631/208/2491/2046
631/449/1659
631/449/2669
Agrobacterium - genetics
Aquatic plants
DNA, Plant - genetics
Efficiency
Flowers & plants
Gene Knockout Techniques
Gene Targeting
Genes
Genetic analysis
Genetic transformation
Genetics
Genomes
Genomics
Genotype
Genotype & phenotype
Homologous Recombination
Humanities and Social Sciences
Integration
Marchantia - genetics
Membrane Proteins - genetics
multidisciplinary
Negative selection
NOP1 gene
Plant Proteins - genetics
Plants, Genetically Modified - genetics
Science
Thalli
Transformation, Genetic
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Summary:The liverwort Marchantia polymorpha is an emerging model organism on account of its ideal characteristics for molecular genetics in addition to occupying a crucial position in the evolution of land plants. Here we describe a method for gene targeting by applying a positive/negative selection system for reduction of non-homologous random integration to an efficient Agrobacterium -mediated transformation system using M. polymorpha sporelings. The targeting efficiency was evaluated by knocking out the NOP1 gene, which impaired air-chamber formation. Homologous recombination was observed in about 2% of the thalli that passed the positive/negative selection. With the advantage of utilizing the haploid gametophytic generation, this strategy should facilitate further molecular genetic analysis of M. polymorpha , in which many of the mechanisms found in land plants are conserved, yet in a less complex form.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep01532